Phellinus igniarius double-layer superimposed substitute culture medium and application thereof

Abstract

This technical solution belongs to the field of edible and medicinal fungi cultivation technology. In order to stabilize production, shorten the cycle, increase yield, and thus effectively realize the year-round industrialized cultivation of Sanghuang and the standardized and regulated production system of the entire green and safe Sanghuang industry chain, and further enhance the level and influence of the Sanghuang industry, this invention provides a double-layer superimposed substrate cultivation medium for Sanghuang and its application. The medium consists of two specific substrates forming a double-layer superimposed medium. The upper substrate components include mulberry wood chips, oak wood chips, cottonseed hulls, wheat bran, white sugar, gypsum, lime, perlite, corn cob, and water. The lower substrate specific components include mulberry wood chips, fruit tree branch wood chips, cottonseed hulls, wheat bran, white sugar, gypsum, lime, corn cob, and water. After application through steps such as seed production, medium preparation, inoculation and cultivation, yellowing management, and harvesting, significant technical effects have been unexpectedly achieved, such as a fresh weight of over 71g / bag, a biological efficiency of over 16%, and an overall cultivation cycle as short as 94 days.

Description

Technical Field

[0001] This invention belongs to the field of edible and medicinal fungi cultivation technology, specifically involving a double-layer superimposed substrate cultivation medium for Phellinus linteus and its application. Background Technology

[0002] Sanghuang belongs to the phylum Basidiomycota, class Agaricomycetes, order Hymenochaetales, family Hymenochaetaceae, and genus Sanghuang. Sanghuangporus It is a general term for a group of fungi, a precious medicinal and edible fungus, known as "forest gold". It has rich nutritional value and contains a variety of flavonoids, polysaccharides, total phenols, terpenes, furans and other substances. It has the effects of immune regulation, anti-tumor, liver protection, anti-oxidation, anti-inflammation and hypoglycemia, and has a broad market prospect.

[0003] Phellinus linteus, a rare edible and medicinal fungus, has been recorded in traditional Chinese medicine texts throughout history, but it did not receive widespread attention or application in my country until the early 1990s. This rare fungal resource began to attract the attention of scholars and industry professionals, and breakthroughs in artificial domestication gradually led to large-scale regional cultivation. Currently, there are two cultivation methods for Phellinus linteus: log cultivation and substrate cultivation. Log cultivation typically requires three years to fully utilize the wood substrate and achieve a high bioconversion rate, while substrate cultivation usually requires one year and allows for one to two harvests per year. The substrate-based cultivation model started in 2010, and in recent years, it has gradually overcome technical challenges in areas such as new variety selection, liquid spawn technology, and factory-scale year-round cultivation. However, the scale of the factory-scale Sanghuang industry using substrate-based cultivation has not expanded significantly. This is mainly because the current factory-scale substrate-based cultivation model still faces many technical problems that hinder the standardized production of the entire green and safe Sanghuang industry chain. These problems include the extensive nature of seasonal outdoor cultivation, high pollution rates, issues with pesticide residue or heavy metal control, slow yellowing, long cultivation cycles or low production efficiency, and low yield or biological efficiency. As a result, the Sanghuang industry has not yet formed a green, efficient, healthy, and sustainable development model.

[0004] Existing technologies for optimizing the substrate cultivation of Sanghuang mainly focus on screening suitable substrate formulas, optimizing inoculation methods, and optimizing mycelial and fruiting body growth conditions. For example, patent number CN201910712740.0, entitled "A Method for Factory-Scale Bottle Cultivation of Sanghuang," uses bottle cultivation with substrate as a substitute. It focuses on a series of measures, including the adoption of formulated raw materials and environmental control, aiming to provide a suitable factory-scale cultivation method to improve production efficiency, shorten the production cycle, and increase the yield and quality of Sanghuang. Ultimately, it achieves technical effects such as shortening the mycelium growth time (dark cultivation or full mycelium growth) to 28-35 days, a biological efficiency of 4.66%, and a yield (fresh weight) of 16.3g / bottle.

[0005] Liu Yan et al. optimized the cultivation conditions for *Sanghuang* using substrate substitutes, aiming to shorten the cultivation cycle, increase the yield and quality of *Sanghuang*, and enhance the added value of mulberry trees. Their work focused on the substrate substitute formula, temperature and humidity during fruiting body production, inoculation methods, and greenhouse sterilization effects, achieving optimal results such as a full-body growth time of 53 days and a fresh weight of 149g / bag. Patent number CN202111006288.X, entitled "A Method for Cultivating *Sanghuang*", also focuses on a series of measures including specific formula raw materials and environmental control, achieving optimal results such as a cultivation cycle of 50-70 days, a cultivation period (including fruiting body management) of over 41 days, and a fresh weight of 31.8g / bag. Lu Chunxia et al.'s study on the effects of different culture medium formulas on the growth of *Sanghuang* mycelium and fruiting bodies revealed a cultivation model requiring over 52 days for full-body growth, over 15 days for primordia formation, and over 57 days for fruiting body maturity, resulting in an overall production cycle as long as 124 days. The aforementioned reports all focus on achieving corresponding technical effects through methods such as raw material formulation and environmental parameter control. However, the results or models either have low yields, small-scale experiments with long overall cycles, or low production efficiency and uncontrollable quality, making them unsuitable for standardized factory production. There are currently no reports on the technical information regarding the combined formulation of two raw material substitutes and their effective application in the cultivation of *Sanghuang* (a type of medicinal mushroom).

[0006] To stabilize production, shorten the cycle, and increase yield, thereby effectively realizing the year-round industrialized cultivation and production of Sanghuang, achieving a standardized and regulated production system for the entire green and safe Sanghuang industrial chain, and further enhancing the level and influence of the Sanghuang industry, this technical solution provides a double-layer superimposed substrate cultivation medium for Sanghuang and its application. This medium can achieve the following technical effects: mycelium can fully grow into the bag in only 18-24 days, and the entire production cycle can be shortened to within 94 days. The Sanghuang mycelium can rapidly differentiate and grow (easily forming Sanghuang primordia) and the yield can be as high as 71g / bag (fresh weight) or more, with a biological efficiency of over 16%. Summary of the Invention

[0007] To stabilize production, shorten the cycle, and increase yield, thereby effectively realizing the year-round industrialized cultivation and production of Sanghuang (a type of medicinal mushroom), and achieving a standardized and regulated production system for the entire green and safe Sanghuang industry chain, further enhancing the level and influence of the Sanghuang industry, this technical solution provides a double-layer superimposed substrate cultivation medium for Sanghuang and its application. Specifically, the medium consists of two layers of specific substrates. The upper substrate components include mulberry sawdust, oak sawdust, cottonseed hulls, wheat bran, white sugar, gypsum, lime, perlite, corn cobs, and water. The lower substrate components include mulberry sawdust, fruit tree branch sawdust, cottonseed hulls, and wheat bran. The substrate consists of: bark, white sugar, gypsum, lime, corn cobs, and water. The upper substrate comprises, by mass ratio: 20-30% mulberry sawdust, 20-30% oak sawdust, 20-30% cottonseed hulls, 5-20% wheat bran, 1-2% white sugar, 1-2% gypsum, 1-2% lime, and 1-2‰ perlite, with a moisture content of 55-60%. The lower substrate comprises, by mass ratio: 20-30% mulberry sawdust, 20-30% fruit tree branch sawdust, 20-30% cottonseed hulls, 5-20% wheat bran, 1-2% white sugar, 1-2% gypsum, 1-2% lime, and 1-2% corn cobs, with a moisture content of 55-60%.

[0008] The steps for applying this cultivation medium method include the following:

[0009] 1. Seed production

[0010] Prepare liquid culture medium, sterilize and cool it for later use; inoculate with Sanghuang fungus in test tubes, and culture at 25-28℃ and 150rpm for 9-10 days to obtain Sanghuang liquid culture.

[0011] 2. Prepare double-layer stacked culture medium

[0012] Weigh the raw materials according to the above base material ratio, mix them evenly, and then pack them into bags. Fill one end of the bag with the upper layer of base material and the other end with the lower layer of base material. After sealing the bag, sterilize it under high pressure and moist heat at 115-120℃ for 240-270 minutes; then cool it for later use.

[0013] 3. Inoculation and Culture

[0014] Inoculate with liquid mulberry fungus, transfer to a culture room for mycelial growth management, and culture in the dark at a temperature of 26-30℃ for 20-24 days until the mycelium completely covers the culture medium, and then perform light color change for 3-5 days;

[0015] 4. Management of Yellowing

[0016] After the mycelium on the culture medium changes color, an opening is made at the junction of the upper and lower substrates. The culture is then maintained for 3-5 days under diffused light conditions of 25-31℃, 80-99% humidity, 500-3000ppm carbon dioxide, and 100-500Lux light intensity to promote rapid knotting and differentiation of the *Sanghuang* mycelium at the opening. After the primordia have fully formed at the opening, the culture is maintained for 20-25 days under diffused light conditions of 26-30℃, 90-99% humidity, 500-2500ppm carbon dioxide, and 100-200Lux light intensity. Finally, the culture is maintained for 20-30 days under diffused light conditions of 27-29℃, 90-99% humidity, 500-2000ppm carbon dioxide, and 100-300Lux light intensity.

[0017] 5. Harvesting

[0018] Before harvesting, stop humidifying for 3-5 days. Harvesting can begin after the fruiting bodies of Phellinus linteus have become leathery.

[0019] Compared with the prior art, the advantages of the present invention are:

[0020] 1. For the first time in this field, the two formulas of substitute materials were combined and effectively applied to the cultivation of Phellinus linteus, unexpectedly achieving significant technical effects such as fresh weight of over 71g / bag and biological efficiency of over 16%.

[0021] 2. The composition and ratio of the culture medium raw materials are reasonable, which leads to faster mycelial growth. Although the two culture media form mycelia or mycelial bodies with two different growth patterns, they can unexpectedly and easily form Phlox sanghuang primordia and then enter reproductive growth after extending, crossing and twisting at the interface. That is, the primordia formation time is short (3-5 days after color change). The changes in the "quality and quantity" of yield have achieved unexpected technical effects, which not only improve the stability of yellowing (production stability) but also shorten the cultivation cycle to a certain extent.

[0022] 3. By combining the biological characteristics of Sanghuang, targeted environmental parameter regulation is carried out to stabilize the production process while promoting growth in a synergistic way, so that Sanghuang fruiting bodies with beautiful shape and high yield can be produced through year-round cultivation.

[0023] 4. The raw materials are simple and easy to obtain, the process is simple and easy to operate, the production cost is low and the production efficiency is high, the cultivation method is scientific and controllable and can be replicated without being affected by the season, the quality, safety and production environment are controllable, and the overall cultivation cycle is as short as 94 days. It is suitable for the standardized, year-round and factory production of green and safe Sanghuang, which can ensure the healthy and sustainable development of the Sanghuang industry. Attached Figure Description

[0024] Figure 1 The production data are for the example and control groups. Implementation Example

[0025] 1. Seed production

[0026] Prepare liquid culture medium, sterilize and cool it for later use; inoculate with Sanghuang fungus in test tubes, and culture at 25-28℃ and 150rpm for 9-10 days to obtain Sanghuang liquid culture.

[0027] 2. Prepare double-layer stacked culture medium

[0028] The upper substrate contains 27% mulberry wood chips, 24.9% oak wood chips, 25% cottonseed hulls, 20% wheat bran, 1% white sugar, 1% gypsum, 1% lime, and 1‰ perlite, with a moisture content of 55-60%. The lower substrate contains 27% mulberry wood chips, 24% fruit tree branches and wood chips, 25% cottonseed hulls, 20% wheat bran, 1% white sugar, 1% gypsum, 1% lime, and 1% corn cob, with a moisture content of 55-60%.

[0029] Weigh the raw materials according to the above base material ratio, mix them evenly, and then bag them. Pack 500g of each material into a bag. Put the upper layer of base material into one end of the bag and the lower layer of base material into the other end. After sealing the bags, sterilize them under high pressure and moist heat at 115-120℃ for 240-270 minutes; then cool them for later use.

[0030] 3. Inoculation and Culture

[0031] Inoculate with liquid mulberry fungus, transfer to the culture room for mycelial growth management, and culture in the dark at 28℃ for 20 days until the mycelium completely covers the culture medium, and then perform light color change for 4 days.

[0032] 4. Management of Yellowing

[0033] After the mycelium on the culture medium changes color, an opening is made at the junction of the upper and lower substrates. The culture is then maintained for 4 days under diffused light conditions of 25-31℃, 80-99% humidity, 500-3000ppm carbon dioxide, and 100-500Lux light intensity to promote rapid knotting and differentiation of the mycelium at the opening. After the primordia have fully formed at the opening, the culture is maintained for 21 days under diffused light conditions of 26-30℃, 90-99% humidity, 500-2500ppm carbon dioxide, and 100-200Lux light intensity, followed by 22 days under diffused light conditions of 27-29℃, 90-99% humidity, 500-2000ppm carbon dioxide, and 100-300Lux light intensity.

[0034] 5. Harvesting

[0035] Before harvesting, stop humidifying for 3 days. Harvesting can begin after the fruiting bodies of Phellinus linteus have become leathery. Example

[0036] 1. Seed production

[0037] Prepare liquid culture medium, sterilize and cool it for later use; inoculate with Sanghuang fungus in test tubes, and culture at 25-28℃ and 150rpm for 9-10 days to obtain Sanghuang liquid culture.

[0038] 2. Prepare double-layer stacked culture medium

[0039] The upper substrate contains 28% mulberry wood chips, 26.9% oak wood chips, 30% cottonseed hulls, 12% wheat bran, 1% white sugar, 1% gypsum, 1% lime, and 1‰ perlite, with a moisture content of 55-60%. The lower substrate contains 28% mulberry wood chips, 26% fruit tree branches and wood chips, 30% cottonseed hulls, 12% wheat bran, 1% white sugar, 1% gypsum, 1% lime, and 1% corn cob, with a moisture content of 55-60%.

[0040] Weigh the raw materials according to the above base material ratio, mix them evenly, and then bag them. Pack 500g of each material into a bag. Put the upper layer of base material into one end of the bag and the lower layer of base material into the other end. After sealing the bags, sterilize them under high pressure and moist heat at 115-120℃ for 240-270 minutes; then cool them for later use.

[0041] 3. Inoculation and Culture

[0042] Inoculate with liquid sanghuang culture, transfer to the culture room for mycelial growth management, and culture in the dark at 28℃ for 20 days until the mycelium completely covers the culture medium, and then perform light color change for 3 days;

[0043] 4. Management of Yellowing

[0044] After the mycelium on the culture medium changes color, an opening is made at the junction of the upper and lower substrates. The culture is then maintained for 4 days under diffused light conditions of 25-31℃, 80-99% humidity, 500-3000ppm carbon dioxide, and 100-500Lux light intensity to promote rapid knotting and differentiation of the *Sanghuang* mycelium at the opening. After the primordia have fully formed at the opening, the culture is maintained for 20 days under diffused light conditions of 26-30℃, 90-99% humidity, 500-2500ppm carbon dioxide, and 100-200Lux light intensity, followed by 25 days of culture under diffused light conditions of 27-29℃, 90-99% humidity, 500-2000ppm carbon dioxide, and 100-300Lux light intensity.

[0045] 5. Harvesting

[0046] Before harvesting, stop humidifying for 3 days. Harvesting can begin after the fruiting bodies of Phellinus linteus have become leathery. Example

[0047] 1. Seed production

[0048] Prepare liquid culture medium, sterilize and cool it for later use; inoculate with Sanghuang fungus in test tubes, and culture at 25-28℃ and 150rpm for 9-10 days to obtain Sanghuang liquid culture.

[0049] 2. Prepare double-layer stacked culture medium

[0050] The upper substrate contains 29% mulberry wood chips, 24.9% oak wood chips, 28% cottonseed hulls, 15% wheat bran, 1% white sugar, 1% gypsum, 1% lime, and 1‰ perlite, with a moisture content of 55-60%. The lower substrate contains 29% mulberry wood chips, 24% fruit tree branches and wood chips, 28% cottonseed hulls, 15% wheat bran, 1% white sugar, 1% gypsum, 1% lime, and 1% corn cob, with a moisture content of 55-60%.

[0051] Weigh the raw materials according to the above base material ratio, mix them evenly, and then bag them. Pack 500g of each material into a bag. Put the upper layer of base material into one end of the bag and the lower layer of base material into the other end. After sealing the bags, sterilize them under high pressure and moist heat at 115-120℃ for 240-270 minutes; then cool them for later use.

[0052] 3. Inoculation and Culture

[0053] Inoculate with liquid mulberry fungus, transfer to the culture room for mycelial growth management, and culture in the dark at 28℃ for 22 days until the mycelium completely covers the culture medium, and then perform light color change for 5 days;

[0054] 4. Management of Yellowing

[0055] After the mycelia on the culture medium change color, an opening is made at the junction of the upper and lower substrates. The culture is then maintained for 4 days under diffused light conditions of 25-31℃, 80-99% humidity, 500-3000ppm carbon dioxide, and 100-500Lux light intensity to promote rapid knotting and differentiation of the mycelia at the opening. After the primordia have fully formed at the opening, the culture is maintained for 21 days under diffused light conditions of 26-30℃, 90-99% humidity, 500-2500ppm carbon dioxide, and 100-200Lux light intensity, followed by 27 days under diffused light conditions of 27-29℃, 90-99% humidity, 500-2000ppm carbon dioxide, and 100-300Lux light intensity.

[0056] 5. Harvesting

[0057] Before harvesting, stop humidifying for 4 days. Harvesting can begin after the fruiting bodies of Phellinus linteus have become leathery.

Claims

1. A double-layer superimposed substrate culture medium for the cultivation of Phellinus linteus, characterized in that, The culture medium consists of two layers of specific substrate. The upper substrate includes mulberry wood chips, oak wood chips, cottonseed hulls, wheat bran, white sugar, gypsum, lime, perlite, corn cob, and water. The lower substrate includes mulberry wood chips, fruit tree branches and wood chips, cottonseed hulls, wheat bran, white sugar, gypsum, lime, corn cob, and water. The upper substrate composition is as follows: 20-30% mulberry wood chips, 20-30% oak wood chips, 20-30% cottonseed hulls, 5-20% wheat bran, 1-2% white sugar, 1-2% gypsum, 1-2% lime, 1-2‰ perlite, and a moisture content of 55-60%. The lower substrate consists of the following components in the following mass ratio: 20-30% mulberry wood chips, 20-30% fruit tree branch wood chips, 20-30% cottonseed hulls, 5-20% wheat bran, 1-2% white sugar, 1-2% gypsum, 1-2% lime, 1-2% corn cob, and a moisture content of 55-60%.

2. A method for applying the double-layer superimposed substrate culture medium for *Sanghuang* as described in claim 1, characterized in that, Includes the following steps: (1) Seed preparation: Prepare liquid culture medium, sterilize and cool it for later use; inoculate with Sanghuang fungus test tube culture, and culture at 25-28℃ and 150rpm for 9-10 days to obtain Sanghuang liquid culture; (2) Preparation of double-layer superimposed culture medium: Weigh the raw materials according to the base material ratio described in claim 1, stir evenly and then put them into bags. Put the upper base material into one end of the bag and the lower base material into the other end of the bag. After sealing the bag, sterilize it under high pressure and moist heat at a temperature of 115-120℃ for 240-270 minutes; cool and wait for use. (3) Inoculation and culture: Inoculate with liquid mulberry fungus, transfer to the culture room for mycelial growth management, and culture in the dark at a temperature of 26-30℃ for 20-24 days until the mycelium completely covers the culture medium, and then perform light color change for 3-5 days; (4) Yellowing management: After the mycelium in the culture medium changes color, open the opening at the junction of the upper and lower substrates and culture it for 3-5 days under the conditions of temperature 25-31℃, humidity 80-99%, carbon dioxide concentration 500-3000ppm, and light intensity 100-500Lux diffused light to promote the rapid twisting and differentiation of the mycelium at the opening; after the primordia at the opening are fully formed, culture it for 20-25 days under the conditions of temperature 26-30℃, humidity 90-99%, carbon dioxide concentration 500-2500ppm, and light intensity 100-200Lux diffused light, and then culture it for 20-30 days under the conditions of temperature 27-29℃, humidity 90-99%, carbon dioxide concentration 500-2000ppm, and light intensity 100-300Lux diffused light. (5) Harvesting: Before harvesting, stop humidifying for 3-5 days. Harvesting begins after the fruiting bodies of Phellinus linteus have become leathery.

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